顽拗植物澳洲坚果成熟叶片DNA提取方法比较

A Comparative Study on Genomic DNA Extraction from Mature Leaves of Recalcitrant Macadamia

目的：针对澳洲坚果成熟叶片中富含多糖、多酚等杂质的特点，建立澳洲坚果成熟叶片中提取高质量DNA的方法。方法：采用改良CTAB法和改良SDS法提取澳洲坚果样品的总DNA,并对产物进行紫外、电泳及PCR扩增检测。结果：改良CTAB法的平均产率为13．6μg／g，略低于改良SDS法18．5μg/g,但改良CTAB法可有效去除多糖等杂质，获得的基因组DNA质量高。OD260／OD280均在1．7—1．9之间。进行LCSR扩增可获得清晰、多态性好的条带。结论：改良CTAB法较之改良SDS法更适合于从澳洲坚果成熟叶片中提取高质量DNA。

Objective： To select the best method to extract DNA from mature leaves of Maeadamia which was rich in polysaceharide and polyphe- nol. Method： The improved CTAB method and improved SDS method were used to extract DNA. And then detect the quality of DNA by UV scanning, eleetrophoresis and PCR amplifieation. Results： The extract ratio of improved CTAB method was higher than improved SDS method. Improved CTAB method could remove polysaeeharides and other impurity and can obtain the pure genetic DNA, the value of OD260/OD280 was 1.7 to 1.9, and then ISSR analysis could produced clear polymorphic patterns. Conclusion： Improved CTAB method was better than improved SDS method to extract high quality DNA from mature leaves of Maeadamia.